Extinction of Cocaine Self-Administration Reveals Functionally and Temporally Distinct Dopaminergic Signals in the Nucleus Accumbens
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Neuron, Vol. 46, 661–669, May 19, 2005, Copyright ©2005 by Elsevier Inc. DOI 10.1016/j.neuron.2005.04.036 Extinction of Cocaine Self-Administration Reveals Functionally and Temporally Distinct Dopaminergic Signals in the Nucleus Accumbens Garret D. Stuber,1 R. Mark Wightman,1,3 2004), which can then become associated with neutral and Regina M. Carelli1,2,* environmental stimuli. Because of this, dopaminergic 1Curriculum in Neurobiology transmission within the NAc may promote the formation 2 Department of Psychology of an unnatural relationship between drugs of abuse 3 Department of Chemistry and Neuroscience Center and environmental cues (O’Brien et al., 1993; Everitt et The University of North Carolina al., 1999; Hyman and Malenka, 2001; Wise, 2004). How- Chapel Hill, North Carolina 27599 ever, because cocaine has a potent pharmacological effect to inhibit monoamine uptake (Jones et al., 1995; Wu et al., 2001), it has been difficult to separate in- Summary creases in dopamine due to either the primary pharma- cological or secondary conditioned effects of the drug. While Pavlovian and operant conditioning influence Microdialysis studies in self-administrating animals drug-seeking behavior, the role of rapid dopamine have shown that NAc dopamine decreases over min- signaling in modulating these processes is unknown. utes leading up to a lever press for cocaine and then During self-administration of cocaine, two dopami- increases slowly after the drug infusion (Wise et al., nergic signals, measured with 100 ms resolution, oc- 1995). When dopamine changes are examined with curred immediately before and after the lever press greater temporal resolution, electrochemical studies (termed pre- and postresponse dopamine transients). have revealed phasic changes in dopamine (lasting Extinction of self-administration revealed that these only a few seconds), which are time locked to the oper- two signals were functionally distinct. Preresponse ant response for the drug (Phillips et al., 2003b; Stuber transients, which could reflect the motivation to ob- et al., 2005). Consistent with this, subsequent exposure tain the drug, did not decline during extinction. Re- to drug-associated cues led to an increase of dopa- markably, postresponse dopamine transients attenu- mine within the NAc (Ito et al., 2000; Phillips et al., ated as extinction progressed, suggesting that they 2003b) and can lead to the relapse of drug taking, thus encode the learned association between environmen- playing an important role in the manifestation and tal cues and cocaine. A third type of dopamine tran- maintenance of addiction (Weiss et al., 2001; Crombag sient, not time locked to overt stimuli, decreased in et al., 2002). frequency during extinction and correlated with cal- While reward-related learning utilizes dopaminergic culated cocaine concentrations. These results show transmission to associate environmental stimuli with a that dopamine release transients involved in different primary reinforcer (Schultz, 1998; Ito et al., 2000), the aspects of cocaine self-administration are highly precise role that dopaminergic signaling plays during plastic—differentially governed by motivation, learned drug self-administration remains unknown. To address associations linked with environmental stimuli, and this, cyclic voltammetry with 100 ms temporal resolu- the pharmacological actions of cocaine. tion was used to monitor rapid dopamine release dur- ing regular cocaine self-administration (maintenance), Introduction extinction, and reinstatement. The superb temporal res- olution of this technique revealed that dopaminergic Dopaminergic neurons originating in the ventral teg- signals play three temporally and functionally distinct mental area and projecting to the amygdala, prefrontal roles that are differentially influenced by extinction. cortex, and nucleus accumbens (NAc) are hypothe- sized to relate information about a primary reinforcer, Results such as cocaine, with goal-directed behavior and/or environmental stimuli (McClure et al., 2003; Schultz, Cocaine Self-Administration during 2004). This idea is supported by electrophysiological Extinction/Reinstatement studies in behaving animals, which have demonstrated Intravenously catheterized rats were initially trained to that dopaminergic neurons are initially activated by pri- self-administer cocaine (0.33 mg/infusion; delivered mary reinforcers but adapt over repeated pairings to over 6 s) in 2 hr daily sessions and were surgically pre- respond to conditioned stimuli that are associated with pared for monitoring rapid dopamine signaling in the the reinforcer (Schultz, 1998). Furthermore, the degree NAc core after 2–3 weeks of training. Following recov- of phasic activity of dopamine neurons is a function of ery from surgery and 3–5 days of retraining, rats un- the probability in which a conditioned stimulus predicts derwent a within-session extinction/reinstatement ex- a subsequent reward (Fiorillo et al., 2003), suggesting periment conducted in three phases. In phase one that dopaminergic transmission can adapt to changing (termed maintenance), rats completed two to four lever degrees of uncertainty of which a cue can predict a press responses in quick succession followed by five reward. lever presses with mean interinfusion intervals of 439 ± Virtually all drugs of abuse cause the release of dopa- 33 s. During maintenance, each lever press resulted in mine in the NAc (Di Chiara and Imperato, 1988; Wise, an intravenous cocaine infusion (0.33 mg, 6 s) paired with drug-associated cues (20 s). Lever pressing beha- *Correspondence: [email protected] vior remained stable throughout maintenance, with no Neuron 662 Figure 1. Lever Press Behavior during Main- tenance, Extinction, and Reinstatement of Cocaine Self-Administration (A) Representative behavioral response rec- ord from one animal throughout the three ex- perimental phases. Each vertical tick mark represents a lever press response. The ani- mal was reinstated to self-administer co- caine after extinction by issuing a noncontin- gent “priming” infusion of cocaine paired with a 20 s presentation of the drug-associ- ated cues (indicated by the arrow). (B–D) Interresponse intervals for lever presses in maintenance, extinction, and reinstatement. significant difference in the interresponse interval over dopamine signaling relates to the association of lever the trials [F(4, 39) = 0.48; p = 0.74; Figures 1A and 1B]. press responding and the cues that predict cocaine de- In phase two (extinction), saline was substituted for co- livery (Phillips et al., 2003b). Therefore, we hypothe- caine. Each lever press resulted in saline infusion (6 s) sized that the preresponse phasic dopamine signal paired with the drug-associated cues (20 s). This led to would remain during extinction, while the postresponse a dramatic increase in the rate of responding measured dopamine signal would attenuate as the learned asso- over the first ten trials of extinction [F(9, 79) = 6.2; p % ciation between the reinforcer and the drug-associated 0.0001; Figures 1A and 1C] and the eventual cessation cues decreased across extinction trials. For analysis of responding after 20.3 ± 3.0 trials. Following 30 min purposes, extinction was broken up into early (first ten) of no responding, phase three (reinstatement) was initi- and late (last five) lever presses. Figure 3A shows a sin- ated. Specifically, self-administration behavior was re- gle trial in early extinction. Even though each lever established by a noncontingent intravenous infusion of press resulted in a saline infusion paired with the drug- cocaine (0.33 mg, 6 s) paired with the same drug-asso- associated cues, both the pre- and postresponses ciated stimuli. During the reinstatement phase, rats were associated with the lever press. A repeated mea- were allowed to respond five times, in which each response resulted in cocaine paired with the drug- sures ANOVA revealed that preresponse DA transients associated cues, as in maintenance. Response rates remained remarkably stable in amplitude throughout decreased and eventually stabilized over the five early extinction and were not significantly different rela- reinstatement trials [F(4, 39) = 3.1; p < 0.05; Figure 1D]. tive to the maintenance phase [F(10, 87) = 0.83; p = 0.60; Figure 3B]. In contrast, the amplitude of postre- Phasic Dopamine Signals Are Associated with the sponse dopamine transients was maintained during Operant Response during Maintenance early extinction (Figures 3A and 3C) but significantly de- Phasic dopamine time locked to a single operant re- clined as extinction proceeded [repeated measures sponse during maintenance is shown in Figure 2A. As ANOVA; F(10, 87) = 2.44; p % 0.05; Figure 3C]. Post hoc reported previously (Phillips et al., 2003b; Stuber et al., comparisons revealed that the postresponse dopamine 2005), dopaminergic signals at the lever press in main- signal on trial 10 was significantly less than the average tenance have two distinct components: an initial in- dopamine signal during the maintenance phase (p % crease in dopamine in the time interval 10 s before the 0.05). lever press response (termed preresponse) followed by As the extinction phase proceeded, a clear dissoci- a larger increase within 10 s after the response (termed ation between pre- and postresponse dopamine tran- postresponse). The mean preresponse dopaminergic sients became apparent. Figure 3D shows a single lever signal was 52.6 ± 6.2 nM and remained stable in ampli- press response (from